Genipin suppression of fibrogenic behaviors of the alpha-TN4 lens epithelial cell line.
Genipin suppression of fibrogenic behaviors of the alpha-TN4 lens epithelial cell line.
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发表时间：
2006
影响因子：
2.8
通讯作者：
A. Kitano;S. Saika;O. Yamanaka;P. Reinach;K. Ikeda;Y. Okada;K. Shirai;Y. Ohnishi
中科院分区：
文献类型：
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作者：
A. Kitano;S. Saika;O. Yamanaka;P. Reinach;K. Ikeda;Y. Okada;K. Shirai;Y. Ohnishi
目的在透镜上皮细胞系中，确定α-TN 4，京尼平，草药inchin-ko-to的肠代谢物成分，是否抑制促纤维化肌成纤维细胞的产生和纤维化细胞因子的上调，并评估药物在预防后囊膜混浊（PCO）中的潜在益处。单位：日本和歌山，和歌山医科大学眼科。方法采用比色法、划痕法、免疫组化法和Western blot法检测肌成纤维细胞生成的标志--α-平滑肌肌动蛋白（α-SMA）的表达和α-TN 4细胞的增殖、迁移。转化生长因子-β 1（TGF-β 1）和结缔组织生长因子（CTGF）的基因表达的特点与实时逆转录聚合酶链反应。此外，通过Western印迹和免疫组织化学评估p38丝裂原活化蛋白激酶（p38 MAPK）、细胞外信号调节激酶（ERK）分支和Smad信号。使用用于培养基中的核基质蛋白41/7（NMP 41/7）的商业比色测定试剂盒评价京尼平的细胞毒性。结果京尼平可抑制细胞增殖和迁移，并抑制Smad和p38 MAPK磷酸化，但ERK信号通路增强。京尼平抑制TGF-β 1和CTGF的mRNA表达。基于较低强度的α-SMA免疫细胞化学染色，细胞质纤维形成下降。然而，α-SMA蛋白表达实际上没有改变。该阴性结果表明京尼平减弱了含α-SMA的细胞骨架的形成。用京尼平处理细胞48小时没有增加NMP 41/7向培养基的释放，表明该化合物没有细胞毒性。结论京尼平可抑制α-TN 4透镜细胞的纤维化行为，对预防PCO有一定的治疗价值。
PURPOSE To determine in a lens epithelial cell line, alpha-TN4, whether genipin, an intestinal metabolite component of the herbal medicine inchin-ko-to, suppresses profibrogenic myofibroblast generation and upregulation of fibrogenic cytokines and to evaluate the potential benefit of the medicine in preventing posterior capsule opacification (PCO). SETTING Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan. METHODS In this study, alpha-TN4 cell proliferation, migration, and expression of alpha-smooth muscle actin (alpha-SMA), the hallmark of myofibroblast generation, were assayed with a colorimetric assay, scratch wound assay, immunohistochemistry, and Western blot analysis. Gene expression of transforming growth factor-beta1 (TGF-beta1) and connective tissue growth factor (CTGF) was characterized with real-time reverse transcription-polymerase chain reaction. In addition, p38 mitogen-activated protein kinase (p 38 MAPK), extracellular signal-regulated kinase (ERK) limb, and Smad signalings were evaluated by Western blotting and immunohistochemistry. Cytotoxicity of genipin was evaluated using a commercial colorimetric assay kit for nuclear matrix protein 41/7 (NMP41/7) in culture medium. RESULTS Genipin suppressed cell proliferation and migration in association with inhibition of Smad and p38 MAPK phosphorylation, although ERK signaling was enhanced. Genipin suppressed mRNA expression of TGF-beta1 and CTGF. Cytoplasmic fiber formation declined based on less intense alpha-SMA immunocytochemical staining. However, alpha-SMA protein expression was actually not altered. This negative result suggests that genipin attenuated formation of alpha-SMA-containing cytoskeleton. Treatment of the cells with genipin for 48 hours did not increase the release of NMP41/7 to the medium, indicating this compound is not cytotoxic. CONCLUSION Because genipin suppressed alpha-TN4 lens cell fibrogenic behaviors, it may be of therapeutic value in preventing PCO.